• Journal of Plant Biotechnology
• /
• 제7권3호
• /
• pp.175-181
• /
• 2005

This work described some essential factors necessary for micro-propagation of banana for mass production of synthetic seeds for germ plasm conservation, and how peroxides activity of conserved tissue was influenced. Shoot tips of field grown plants were used to obtain shoot clusters on shoot proliferation medium (MS medium supplemented with 5 mg/l BAP). Using longitudinally-split shoot tip technique, 18720, 8640, 7488, 2016 plantlets were obtained from one shoot tip of Maghraby, Grand Naine, Balady, and Williams, respectively, in six subculture, one month each, on solid medium. Shoot tips excised from in vitro grown plantlets were encapsulated in calcium-alginate beads and stored at $4^{\circ}C$ for one month on half-strength MS basal medium without growth regulators or sugars. After one month all the viable-conserved synseeds formed shoots when they were transferred to MS basal medium, some of them showed synchronous formation of shoot and root systems in one week. Plants retrieved from encapsulated shoot tips were hardened off and transferred to soil.

• Plant Resources
• /
• 제6권2호
• /
• pp.108-113
• /
• 2003

This study was carried out to obtain basic information for possibility of oral vaccine in carrot using Agrobacteruim -mediated transformation system. The epitope region of porcine epidemic diarrhea virus (PEDV) spike gene which is classified as a member of the Coronaviridae and causes an acute enteritis in pigs was successfully expressed in carrot (Daucus carota) using the Agrobacterium-mediated transformation system. Hypocotyl segments of in vitro germinated plantlets were infected with Agrobacteriun tumefaciens LBA 4404 harboring PEDV spike gene. Embryogenic callus (EC) was induced on MS selection medium with 1 mg/L 2,4-D, 50 mg/L kanamycin and 300 mg/L cefotaxime after 45 days of culture. Subcultured ECs on MS selection medium without 2,4-D were converted to somatic embryos (SE) of various stage; globular, heart and torpedo stage. Putative transgenic embryos were selected on MS medium with 50 mg/L kanamycin and 300 mg/L cefotaxime. Regenerated plantlets from transformed SE were induced on MS medium containing 50 mg/L kanamycin after 30 days of culture. Genomic PCR confirmed the integration of PEDV spike gene into nuclear genome of carrot and northern blot analysis demonstrated the expression of PEDV spike gene in transgenic carrot.

• Journal of Applied Biological Chemistry
• /
• 제42권3호
• /
• pp.113-118
• /
• 1999

The transgenic plant of Citrus species (Citrus aurantium L.) was constructed with a fatty acid desaturase gene using microprojectile bombardment transformation system. The DNA of a fatty acid desaturase gene, fad7, constructed in pBI121 was coated onto tungsten particles ($1.1{\mu}m$) and introduced into callus cells by bombarding with 1100 psi of helium pressure, 1/4 in of gap distance, 7.0 cm of target distance and 27 in Hg of chamber vacuum. The bombarded cells were selected on the medium containing kanamycin. The selected cells were successfully regenerated into plantlets via somatic embryogenesis on the media containing plant growth regulators. The results of polymerase chain reaction analysis of genomic DNAs from the putative transformants showed that the introduced DNAs of fad7 were present in both the selected callus cells and the regenerated plantlets.

• Plant Resources
• /
• 제8권2호
• /
• pp.81-86
• /
• 2005

An efficient plant regeneration system of Gentiana scabra through somatic embryogenesis was established. Leaves and roots of seedlings of Gentiana scabra excised after germination were cultured on MS basal medium with 2,4-D, NAA or BA. Embryogenic callus was obtained on MS medium with 0.5 mg/L 2,4-D alone or 0.1 mg/L 2,4-D combimation with 1.0 mg/L BA after 45 days of culture. These embryogenic calli gave rise to somatic embryos, which subsequently developed into plantlets on MS medium without PGRs. Also, shoots were effectively differentiated from embryogenic callus when root segments were cultured on MS medium supplement with 0.1 mg/L 2,4-D and 1.0 mg/L BA. Shoots were effectively rooted on MS medium without PGRs. In vitro flowers were formed from plantlets cultured on MS medium with $5\%$ sucrose after 60 days of culture.

• 한국자원식물학회지
• /
• 제22권6호
• /
• pp.506-512
• /
• 2009

To study the effect of genotype of donor plants on anther culture, anthers of nine hybrid cultivars (Derby, Special, Bossanova, Minipaprika, Fiesta, Boogie, Phenlene, Kufrah, and Clarity) of sweet pepper (Capsicum annuum L.) were cultured in a petridish containing C medium (Sibi, Dumas De Vaulx medium) supplemented with 0.1 mg/L 2,4-D and 0.1 mg/L kinetin, 3% sucrose and 0.32% Phytagel. The cultures were incubated in the dark at $35^{\circ}C$ for seven days, and then cultured at $25^{\circ}C$ with a photoperiod of 16 hr. daylength for 40 days. Frequency of callus formation and plant regeneration was varied among cultivars. Callus formation was ranged from 6% in Phenlene to 69.8% in Kufrah. The highest percentage of regenerated plantlets was obtained in cv. Phenlene (2.67%) followed by Bossanova (2.41%). Result of ploidy analysis; chromosome number observation and flowcytometry analysis, showed that haploid plants could be developed from all of these hybrid cultivars except cv. Fiesta, where highest percentage of haploid plants were obtained in Minipaprika (40%) followed by cv. Bossanova (36.1%). Haploid plants derived from these hybrid cultivars contained single set of chromosome (12 in numbers), higher stomata density (numbers), and smaller sized stomata as compare to diploid plants. The mean length of stomata was 26.9 ${\mu}m$ in haploid plants and 35.7 ${\mu}m$ in diploids.

• 식물조직배양학회지
• /
• 제28권4호
• /
• pp.185-188
• /
• 2001

Spathiphyllum을 기내에서 증식시킨 후, 액체배지를 첨가하여 신초의 생장 및 발근을 촉진시키고, 기외에서 순화율을 향상시켜 생산비를 감소시키기 위하여 일련의 실험을 실시하였다. 3∼4개의 신초를 가지고 있는 Spathiphyllum floribundum 'Cupid' 의 신초 cluster 절편체를 BA 2.0 mg/L가 첨가된 LS배지에서 8주간 증식시킨 후에 15 mL의 액체배지를 동일용기에 첨가하였다. 액체배지 첨가는 1/2 MS 다량요소+sucrose 50 g/L +활성탄 5.0∼10.0 g/L가 첨가된 배지 15 mL를 첨가하는 것이 식물체의 기내생장 및 발근에 양호하였으며, 온실에서 perlite와 vermiculite가 1 : 1로 혼합된 용토에 발근된 식물체를 재식하면 95% 이상 생존하였다.

• 식물조직배양학회지
• /
• 제21권5호
• /
• pp.299-302
• /
• 1994

0-4mm 크기의 미성숙접합자배를 0.5-8 mg/L 2,4-D가 첨가된 MS배지에서 배양하였다. 87%의 미성숙접합자배가 정단부위로부터 캘러스의 발생없이 체세포배를 형성하였다. 1-2개의 엽원기와 생장점을 포함하는 정단분열절편을 배양하여 체세포배의 발생위치는 유아임을 확인할 수 있었다. 2,4-D 농도가 4 mg/L 이상일 때에는 유아에서 비배발생캘러스만 나타났으나, 60%의 미성숙접합자배가 자엽가장자리에서 체세포배를 형성하였다. 체세포배는 MS 기본배지에서 발아하여 15개의 체세포 중에서 2개가 식물체로 재분화되었다. 재분화된 식물체는 화분에 옮겨서 생육상에서 성숙개체로 발달하였다.

• 한국약용작물학회지
• /
• 제17권3호
• /
• pp.192-197
• /
• 2009

Camptotheca acuminata, a native of South China is a well known natural source of monoterpene-indole alkaloid camptothecin(CPT), one of the most promising anti-tumoural compounds. This study was conducted to optimize plant growth regulators and culture conditions on plantlets regeneration through organogenesis from callus of Camptotheca acuminta. Callus were induced from various explants of in vitro germinated plantlets of C. acuminta using WPM medium containing 0.2 ㎎/L 2,4-D. Hypocotyl segments were exhibited higher embryogenic callus than the other explants. Shoot buds formation from embryogenic callus was affected by plant growth regulators, pre-treated dark condition and liquid culture. Organogenesis was optimal in WPM liquid medium containing 0.5 ㎎/L BA. The dark pre-treatment for 2 weeks before the solid culture was effective for organogenesis. The regenerated shoots were rooted in WPM medium with 0.2 ㎎/L NAA and successfully acclimated in green-house conditions.

• 한국육종학회지
• /
• 제42권5호
• /
• pp.431-438
• /
• 2010

Anthers of sweet pepper $F_1$ cultivar 'Special' were cultured on Dumas De Vaulx (C medium), supplemented with $0.1mgL^{-1}$ 2, 4-D and $0.1mg{\cdot}L^{-1}$ kinetin with 3% sucrose, and 0.32% phytagel. The calluses obtained were further sub-cultured on Murashige and Skoog (MS) medium without growth regulators for regeneration. Regenerated plantlets were grown in plastic pots under plastic house and characterized their cytological and morphological characters in spring, 2008. Twenty percent plantlets were identified as haploid plants after chromosome and ploidy analysis. Haploid plants contained 12 chromosomes, high stomatal density with small stomatal length as compared to diploid plants. Stomatal length in haploids was 23.3% smaller than diploids. Haploid plants were characterized as small leaf and petiole size, poor vigor, thin stem and short plant height, short internodes and small flower buds, fruit size and fruit weight as compared to diploid plants and most of the haploid fruits were seedless. SP55, SP62, SP68, SP72 and SP77 are found high yielding double haploids with high total soluble content (8.6, 8.7, 9.2, 9.1 and $9.8^{\circ}Brix$, respectively) and desirable fruit shape, and recommended them to exploit as inbred lines for heterosis breeding.

• Journal of Plant Biotechnology
• /
• 제46권1호
• /
• pp.17-21
• /
• 2019

Ginger is an important monocotyledonous plant belonging to the family Zingiberaceae. The objective of this study was to investigate the regeneration potential of ginger using leaf base explants. Auxins such as 2, 4-D and NAA in combination with BA were used for initiation of callus. Different combinations of both ammonium ($NH^{4+}$) and nitrate ($NO^{3-}$) were also studied for efficient callus production. High frequency of white friable calli was observed on modified Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 2, 4-D, 0.5 mg/L NAA and 0.5 mg/L BA. The highest shoot induction (92.33%), shootlets number ($7.33{\pm}0.33$) and length ($88.33{\pm}4.40$) mm were achieved on MS media containing 0.5 mg/L BA. Regenerated shoots were transferred to in vitro rooting media containing 1.0 mg/L IBA. Afterwards, plantlets with well-developed root and shoot system were subjected to a twostep hardening process. 71% of plantlets survived after secondary hardening without any abnormal morphology.